Natural combination hormone replacement formulations and therapies

ABSTRACT

Estrogen and progesterone replacement therapies are provided herein. Among others, the following formulations are provided herein: solubilized estradiol without progesterone; micronized progesterone without estradiol; micronized progesterone with partially solubilized progesterone; solubilized estradiol with micronized progesterone; solubilized estradiol with micronized progesterone in combination with partially solubilized progesterone; and solubilized estradiol with solubilized progesterone.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a non-provisional application of and claims priorityto the following U.S. Provisional Patent Applications: U.S. ProvisionalApplication Ser. No. 61/563,408, entitled “NATURAL COMBINATION HORMONEREPLACEMENT THERAPIES” which was filed on Nov. 23, 2011; U.S.Provisional Application Ser. No. 61/661,302, entitled “ESTRADIOLFORMULATIONS” which was filed on Jun. 18, 2012; and U.S. ProvisionalApplication Ser. No. 61/662,265, entitled “PROGESTERONE FORMULATIONS”which was filed on Jun. 20, 2012. All aforementioned applications arehereby incorporated by reference herein in their entirety.

BACKGROUND

1. Field

This disclosure relates to natural estrogen and progesterone replacementtherapies, with formulations provided for each estradiol andprogesterone alone and in combination for the treatment of pre,peri-menopausal, menopausal and post-menopausal females in relation tothe treatment of Estrogen- and Progesterone-deficient States, each asherein below defined.

2. Discussion of the Related Art

Hormone replacement therapy (HRT) is a medical treatment that involvesthe use of one or more of a group of medications designed to increasehormone levels in women who lack adequate hormone production. HRT canmitigate and prevent symptoms caused by diminished circulating estrogenand progesterone hormones regardless as to whether the subject ispre-menopausal, peri-menopausal, menopausal or post-menopausal. However,specific disease states can exist during each stage of menopausalprogression.

HRT is presently available in various forms. One therapy involvesadministration of low dosages of one or more estrogens. Another involvesadministration of progesterone or a chemical analogue, called aprogestin. Progesterone administration acts, among treating otherdisease states, to mitigate certain undesirable side effects fromestrogen administration including, for example, endometrial hyperplasia(thickening), reducing the incidence of endometrial cancer.

Timing for dosage administration is often varied cyclically, withestrogens taken daily and progesterone taken for approximately two weeksof every month; a method often referred to as “Cyclic-Sequential” or“Sequentially-Combined HRT.” This method is intended to mimic thenatural menstrual cycle and typically causes menstruation similar to aperiod after the progesterone is stopped. This regimen is most typicallyused in peri-menopausal or newly menopausal women as the alternativecontinuous method often results in irregular bleeding in such women. Analternate method, a constant dosage with both estrogen and progesteronetaken daily, is called “continuous-combined HRT.” This method usuallyresults in no menstruation and is used most often after a woman has beenmenopausal for some time.

Estrogen, in its various forms, and progesterone, in its various forms,are used in HRT via a variety of administered dosage forms including,for example, via tablets, capsules and patches.

“Bio-identical” hormones, which are identical in chemical structure tothe hormones naturally produced by human bodies can be used and areoften referred to as natural hormone replacement therapy, or NHRT.

These natural or bio-identical hormones are formulated from variousingredients to match the chemical structure and effect of estradiol,estrone, or estriol (the 3 primary estrogens) as well as progesteronethat occur naturally in the human body (endogenous).

Currently, bio-identical estradiol is available in both branded andgeneric FDA approved versions. FDA-approved bio-identical progesteronefor HRT is available as the branded stand-alone drug commerciallyidentified as PROMETRIUM (progesterone, USP) (Abbott Laboratories,Abbott Park, Ill.), with a generic authorized by the innovator, andgeneric products provided by Teva (Israel) and Sofgen Americas, Inc (NewYork). Other products such as PREMPRO (conjugatedestrogens/medroxyprogesterone acetate tablets) and PREMPHASE (conjugatedestrogens plus medroxyprogesterone acetate tablets) (Wyeth Laboratories,a division Pfizer, Inc., New York) provide both continuous-combined andcyclic-sequential products containing PREMARIN (conjugated estrogenstablets) (estrogen derived from mare's urine) and syntheticmedroxyprogesterone acetate. Other products are available. However, noFDA approved product exists on the market today with combinationbio-identical estradiol and bio-identical progesterone.

SUMMARY

According to various embodiments of the disclosure, natural hormonereplacement therapies are provided comprising cyclic/sequential andcontinuous-combined delivery via pharmaceutical formulations ofsolubilized estradiol and micronized and/or partially or completelysolubilized progesterone. Estradiol and micronized and/or partially orcompletely solubilized progesterone delivered together daily can becombined in either a single unit dose or in separate unit doses,typically in a soft capsule. A 28-day or monthly regimen of tablets orcapsules can be packaged in a single blister pack having delivery daysidentified to improve compliance. Various examples formulations ofnatural hormones, and the use of these formulations for hormonereplacement therapies, each in accordance with the invention are setforth below.

BRIEF DESCRIPTION OF THE DRAWINGS/FIGURES

The accompanying drawings, which are incorporated herein and form a partof the specification, illustrate the present disclosure and, togetherwith the description, further serve to explain the principles of thedisclosure and to enable a person skilled in the pertinent art to makeand use the disclosed embodiments.

FIG. 1 illustrates an exemplary manufacturing process of a fill materialin accordance with various embodiments;

FIG. 2 illustrates an exemplary manufacturing process of a softgelmaterial in accordance with various embodiments;

FIG. 3 illustrates an exemplary manufacturing process in accordance withvarious embodiments; and

FIG. 4 illustrates a graph of the particle distribution obtained inExample 10.

DETAILED DESCRIPTION OF THE ILLUSTRATED EMBODIMENTS

Frequently, higher recommended oral dosages of pharmaceuticals arenecessary to treat a given disease state because many active ingredientsare not completely absorbed by a patient in need of treatment. In otherwords, a better-absorbed dosage form of a medicament such as, forexample, progesterone, or dosage forms that provide greater consistencyof absorption of progesterone among subjects, alone or in combinationwith estradiol, may be able to be administered at dosage strengths lowerthan presently recommended, potentially resulting in a reduced orminimized side effect profile, among other potential benefits.

DEFINITIONS

The term “micronized progesterone,” as used herein, includes micronizedprogesterone having an X50 particle size value below about 15 micronsand/or having an X90 particle size value below about 25 microns.

The term “X50,” as used herein, means that one-half of the particles ina sample are smaller in diameter than a given number. For example,micronized progesterone having an X50 of 5 microns means that, for agiven sample of micronized progesterone, one-half of the particles havea diameter of less than 5 microns. Similarly, the term “X90” means thatninety percent (90%) of the particles in a sample are smaller indiameter than a given number.

The term “medium chain,” as used herein means any medium chaincarbon-containing substance, including C4-C18, and including C6-C12substances, fatty acid esters of glycerol, fatty acids, and mono-, di-,and tri-glycerides of such substances.

The term “uniform distribution” means at least one of uniformdispersion, solubility, or lack of agglomeration of progesterone in adissolution test compared to PROMETRIUM (progesterone, USP) at a similardosage strength and the same USP dissolution apparatus.

The term “bioavailability,” as used herein means the concentration of anactive ingredient (e.g., progesterone or estradiol or estrone) in theblood (serum or plasma). The relative bioavailability may be measured asthe concentration in the blood (serum or plasma) versus time. Otherpharmacokinetic (pK) indicators may be used to measure and assessbioavailability, determined by suitable metrics including AUC, C_(max),and optionally, Tmax.

The term “AUC,” as used herein, refers to the area under the curve thatrepresents changes in blood concentration of progesterone, estradiol orestrone over time.

The term, “C_(max)” as used herein, refers to the maximum value of bloodconcentration shown on the curve that represents changes in bloodconcentrations of progesterone, estradiol or estrone over time.

The term, “T_(max)” as used herein, refers to the time that it takes forprogesterone, estradiol or estrone blood concentration to reach themaximum value.

Collectively AUC, C_(max) and, optionally, T_(max) are the principlepharmacokinetic parameters that can characterize the pharmacokineticresponses of a particular drug product such as progesterone in an animalor human subject.

The term “solubilizer,” as used herein, means any substance or mixtureof substances that may be used to enhance the solubility of estradiol,including, for example and without limitation, appropriatepharmaceutically acceptable excipients, such as solvents, co-solvents,surfactants, emulsifiers, oils and carriers.

The term “excipients,” as used herein, refer to non-activepharmaceutical ingredients (“API”) substances such as carriers,solvents, oils, lubricants and others used in formulating pharmaceuticalproducts. They are generally safe for administering to animals,including humans, according to established governmental standards,including those promulgated by the United States Food and DrugAdministration.

The term “oil” as used herein may be any pharmaceutically acceptablesubstance, other than peanut oil, that would suspend and/or solubilizeany suitable progesterone, starting material, or precursor, includingmicronized progesterone as described herein. More specifically, oils mayinclude, for example and without limitation, medium chain fatty acids,generally of the group known as medium chain fatty acids consisting ofat least one mono-, di-, and triglyceride, or derivatives thereof, orcombinations thereof.

“Fully solubilized progesterone” as used herein means progesterone whichis about 100% in solution.

“Partially solubilized progesterone” as used herein means progesteronewhich is in any state of solubilization up to but not including about100%.

DESCRIPTION

Provided herein are the following formulations: solubilized estradiolwithout progesterone; micronized progesterone without estradiol;micronized progesterone with partially solubilized progesterone;solubilized estradiol with micronized progesterone; solubilizedestradiol with micronized progesterone in combination with partiallysolubilized progesterone; and solubilized estradiol with solubilizedprogesterone. The underlying formulation concepts provided herein may beused with other natural or synthetic forms of estradiol andprogesterone. Micronization specifications, aspects and embodiments arefurther defined herein.

Generally, the pharmaceutical formulations described herein are preparedand administered as filled capsules, typically soft capsules of one ormore materials well known in the art including, for example and withoutlimitation, soft gelatin capsules. Micronized progesterone, as describedherein, may also be prepared for administration in tablets or otherwell-known orally administered dosage forms using standard techniques.

Another aspect of the present disclosure includes a pharmaceuticalformulation of micronized progesterone, micronized progesterone withpartially solubilized progesterone and fully solubilized progesterone,wherein said formulation may provide increased progesteronebioavailability in a treated subject compared to the bioavailabilityprovided by PROMETRIUM (progesterone, USP) when administered at equaldosage strengths.

In accordance with various aspects and embodiments, the solubilityproportion (i.e., the proportion of a solute that enters solution) isnotable. The weight ratio of estradiol to the weight of the entiresolution is also notable due to the intended dose amounts, discussedherein. In particular, it is desirable to obtain a target dosage ofestradiol in an amount of solution that may be readily administered viaa capsule. For example, if it is desired to have a dose of estradiol ina capsule of between about 0.125 mg to about 2 mg, it would also bedesirable to have a total solution weight to be between about 250 mg toabout 400 mg, preferably about 300 mg to about 350 mg and morepreferably about 325 mg. In various embodiments, the following weightratios of estradiol to total solution is from about 0.125/50 mg to about0.125/1000 mg, from about 1 mg:500 mg to about 1 mg:50 mg; from about 1mg:250 mg to about 1 mg:60 mg; from about 1 mg:100 mg to about 1 mg:66mg; from about 2 mg/50 mg to about 2 mg/1000 mg. In various embodiments,the target for single dose product is 325 mg, and a target fill weightfor a combination product (e.g., two or more sterol APIs) is 650 mg.

Other aspects of the present disclosure further provide: more uniformdissolution of progesterone, and reduced intra- and inter-patient bloodlevel variability in formulations of progesterone of the presentdisclosure, typically in combinations with solubilized estradiol, whencompared to equal dosages of PROMETRIUM (progesterone, USP). Blood levelvariability is also compared at equal sampling times followingadministration. Not to be limited by theory, these aspects are believedto be influenced by the percentage of solubilized progesterone in arespective formulation wherein such more uniform dissolution ofprogesterone, and lower intra- and inter-patient blood levelvariability, are influenced by a greater proportion of solubilizedprogesterone relative to total progesterone. A reduced food effect withthe present formulations comprising progesterone may also be implicated.

More uniform dissolution of progesterone in a formulation of the presentdisclosure compared to the dissolution of PROMETRIUM (progesterone, USP)at equal dosage strengths and using the same USP apparatus can bedetermined using standard techniques established for API dissolutiontesting, including that which is described in the examples below.

Reduced intra- and inter-patient variability of progesterone formulatedpursuant to the present disclosure compared to PROMETRIUM (progesterone,USP) can be demonstrated via a fed bio-study such as that describedbelow.

Other aspects of the present disclosure includes the use of formulationsas described herein wherein progesterone is at least one API in saidformulation for the treatment of an animal, including humans: forendometrial hyperplasia; for secondary amenorrhea; as a method oftreatment for preterm birth, when said animal has a shortened cervix,and other disease states or conditions treated with supplementalprogesterone (collectively, “Progesterone-deficient States”); and theuse of formulations as described herein wherein estradiol is at leastone API in said formulation for the treatment of an animal, includinghumans, having menopause-related symptoms including, for example,vasomotor symptoms; in relation to treatment of hypoestrogenism relatedsymptoms including, for example and without limitation, hot flashes andnight sweats (vasomotor symptoms), sleep disturbances, mood changes andvulvo-vaginal atrophy; and osteoporosis and other non-menopausal diseasestates or conditions treated with supplemental estrogen. (collectively,“Estrogen-deficient States”), each in a subject in need of treatment,and each with a non-toxic effective amount of said formulations. As usedherein, the term “treatment”, or a derivative thereof, contemplatespartial or complete inhibition of the stated disease state when aformulation as described herein is administered prophylactically orfollowing the onset of the disease state for which such formulation isadministered. For the purposes of the present disclosure, “prophylaxis”refers to administration of the active ingredient(s) to an animal toprotect the animal from any of the disorders set forth herein, as wellas others.

Unless otherwise specified, “natural,” as used herein with reference tohormones discussed herein, means bio-identical hormones formulated tomatch the chemical structure and effect of those that occur naturally inthe human body (endogenous). An exemplary natural estrogen is estradiol(also described as 17β-estradiol and E2) and a natural progestin isprogesterone. An exemplary cyclic/sequential regimen comprises deliveryof from about 0.125 mg to about 2.0 mg of estradiol daily for 14-18days, followed by delivery of from about 0.125 mg to about 2 mg ofestradiol and about 25 mg to about 200 mg of progesterone daily for10-14 days. Cyclic/sequential regimens may be especially useful formenopausal females. Other exemplary dosage strengths for estradiol foruse in the formulations described herein include, without limitation,0.125, 0.25, 0.375, 0.50, 0.625, 0.75, 1.00, 1.125, 1.25, 1.375, 1.50,1.625, 1.75 and 2.00 mg. Other exemplary dosage strengths forprogesterone for use in the formulations described herein include,without limitation, 25, 50, 75, 100, 125, 150, 175, 200 mg, 250 mg, 300mg, 350 mg and 400 mg. These dosage strengths for each of estradiol andprogesterone can be administered in formulations described herein eitheralone or in combination.

Progesterone active pharmaceutical ingredient may be micronized via anyone of the multiple methods typically utilized by the ordinarily skilledartisan. In various embodiments, micronized progesterone has an X50particle size value of less than about 15 microns, less than about 10microns, less than about 5 microns and/or less than about 3 microns. Invarious embodiments, micronized progesterone has an X90 particle sizevalue of less than about 25 microns, less than about 20 microns, and/orless than about 15 microns.

Particle size may be determined in any suitable manner. For example, aBeckman Coulter LS 13 320 Laser Diffraction Particle Size Analyzer (the“Beckman Device”) may be used to determine particle size. As describedabove, particle size may be represented by various metrics, for example,through an X50 particle size, and/or X90 particle size, or similardescriptions of particle size.

The Beckman Device may be used with various modules for introducing asample for analysis. The Beckman Device may be used with the LS 13 320Universal Liquid Module (“ULM”). The ULM is capable of suspendingsamples in the size range of 0.017 μm to 2000 μm. The ULM is a liquidbased module that allows for delivery of the sample to the sensing zone.The ULM recirculates the sample through the Beckman Device. The ULMcomprises two hoses, one for fluid delivery and another for waste. Thetotal volume used may be 125 mL or less. A sample mass of from about 1mg to about 10 g may be used. The ULM may interact with the BeckmanDevice via pins that fit into slots on the ULM. The ULM may use avariety of suspension fluids, for example, water, butonol, ethanol,chloroform, heptanes, toluene, propanol, COULTER Type 1B Dispersant(“Coulter 1B”), and a variety of other suspension fluids. Surfactantsmay also be used, though pump speed should be adjusted to preventexcessive bubbling. Coulter 1B may comprise one or more of acetaldehyde,ethylene oxide, and/or 1,4-dioxane. The Beckman Device may be configuredto use a variety of optical theories, including the Fraunhofer opticalmodel and the Mie Theory.

The Beckman Device may comprise software to control the Beckman Devicewhile the ULM is in use. The software may control, for example, pumpspeed, use of de-bubble routine, rinse routine, sonicate routine, andfill routine, among others. Parameters regarding the sample run may alsobe configured. For example, run length may be set. Though any suitablerun length may be used, in various embodiments, a time period of 30seconds to 120 seconds, and preferably between 30 seconds and 90 secondsmay be used.

The Beckman Device may be used with the LS 13 320 Micro Liquid Module(“MLM”). The MLM is capable of suspending samples in the size range of0.4 μm to 2000 μm. The MLM is a liquid based module that allows fordelivery of the sample to the sensing zone. The MLM includes a stirrer.The total volume used may be 12 mL or less. The MLM may use a variety ofsuspension fluids, both aqueous and non-aqueous.

Each of estradiol and progesterone as described herein can be formulatedalone pursuant to the teachings below. These formulations can beprepared for oral administration or can be combined, based oncompatibility, for co-administration of estradiol and progesterone in asingle oral unit dosage form.

Progesterone formulations of the present disclosure are prepared viablending with a pharmaceutically acceptable oil; generally, the oilcomprises at least one medium chain fatty acid such as medium chainfatty acids consisting of at least one mono-, di-, or triglyceride, orderivatives thereof, or combinations thereof. Optionally added are otherexcipients including, for example and without limitation, anti-oxidants,lubricants and the like. Sufficient oil is used to form a suspension ofmicronized progesterone or, in the alternative, solubilize progesterone.

Pharmaceutically acceptable oils include, without limitation, the use ofat least one of a caproic fatty acid; a caprylic fatty acid; a capricfatty acid; a tauric acid; a myristic acid; a linoleic acid; a succinicacid; a glycerin; mono-, di-, or triglycerides and combinations andderivatives thereof; a polyethylene glycol; a polyethylene glycolglyceride (GELUCIRE (a polyethylene glycol glyceride); GATTEFOSSE SAS,Saint-Priest, France); a propylene glycol; a caprylic/caprictriglyceride (MIGLYOL (caprylic/capric triglyceride); SASOL GermanyGMBH, Hamburg; MIGLYOL (caprylic/capric triglyceride) includes MIGLYOL810 (Caprylic/Capric Triglyceride), MIGLYOL 812 (Caprylic/CapricTriglyceride), MIGLYOL 816 (Caprylic/Capric Triglyceride) and MIGLYOL829 (Caprylic/Capric/Succinic Triglyceride); acaproic/caprylic/capric/lauric triglyceride; a caprylic/capric/linoleictriglyceride; a caprylic/capric/succinic triglyceride; a propyleneglycol monocaprylate; propylene glycol monocaprate; (CAPMUL PG-8(Propylene Glycol Monocaprylate) and CAPMUL PG-10 (Propylene GlycolMonocaprate); the CAPMUL brands are owned by ABITEC, Columbus Ohio); apropylene glycol dicaprylate; a propylene glycol dicaprylate; mediumchain mono- and di-glycerides (CAPMUL MCM (Medium Chain Mono- andDiglycerides)); a diethylene glycol mono ester (including2-(2-Ethoxyethoxy)ethanol: TRANSCUTOL (diethylene glycol mono ester)); adiethylene glycol monoethyl; esters of saturated coconut and palm kerneloil and derivatives thereof; triglycerides of fractionated vegetablefatty acids, and combinations and derivatives thereof.

In other aspects and embodiments, progesterone is fully solubilizedusing, for example and without limitation, sufficient amounts of:TRANSCUTOL (Diethylene glycol monoethyl ether) and MIGLYOL(caprylic/capric triglyceride); TRANSCUTOL (Diethylene glycol monoethylether), MIGLYOL (caprylic/capric triglyceride) and CAPMUL PG-8(Propylene Glycol Monocaprylate) and/or CAPMUL PG-10 (Propylene GlycolMonocaprate); CAPMUL MCM (Medium Chain Mono- and Diglycerides); CAPMULMCM (Medium Chain Mono- and Diglycerides) and a non-ionic surfactant;and CAPMUL MCM (Medium Chain Mono- and Diglycerides) and GELUCIRE (apolyethylene glycol glyceride).

Various ratios of these oils can be used for full solubilization ofprogesterone. CAPMUL MCM (Medium Chain Mono- and Diglycerides) and anon-ionic surfactant can be used at ratios including, for example andwithout limitation: 65:35, 70:30, 75:25, 80:20, 85:15 and 90:10. CAPMULMCM (Medium Chain Mono- and Diglycerides) and GELUCIRE (a polyethyleneglycol glyceride) can be used at ratios including, for example andwithout limitation, 6:4, 7:3, 8:2, and 9:1. Among other combinations,these oils and/or solubilizers, as defined herein, and combinationsthereof, can be used to form combination estradiol and progesteroneformulations of the present disclosure.

Combinations of these oils can produce partially solubilizedprogesterone, depending upon the desired unit dosage amount ofprogesterone. The greater the amount of progesterone per unit dosageform, the less progesterone may be solubilized. The upward limit ofdosage strength per unit dose it generally limited only by the practicalsize of the final dosage form.

In various embodiments, estradiol is partially, substantially orcompletely solubilized. Solubilized estradiol may include estradiol thatis approximately: 90% soluble in a solvent; 93% soluble in a solvent;95% soluble in a solvent; 97% soluble in a solvent; 99% soluble in asolvent; and 100% soluble in a solvent. Solubility may be expressed as amass fraction (% w/w).

In various embodiments, the solubilizing agent is selected from at leastone of a solvent or co-solvent. Suitable solvents and co-solventsinclude any mono-, di- or triglyceride and glycols, and combinationsthereof.

In addition to the oils referenced above for progesterone, which canalso be used as solubilizers for estradiol, other solubilizers include,for example and without limitation, glyceryl mono- and di-caprylates,propylene glycol and 1,2,3-propanetriol (glycerol, glycerin, glycerine).

Anionic and/or non-ionic surfactants can be used in other embodiments ofthe presently disclosed formulations containing estradiol, progesteroneor a combination thereof. In certain embodiments, a non-ionic surfactantis used. Exemplary non-ionic surfactants may include, for example andwithout limitation, one or more of oleic acid, linoleic acid, palmiticacid, and stearic acid. In further embodiments, the non-ionic surfactantmay comprise polyethylene sorbitol esters, including polysorbate 80,which is commercially available under the trademark TWEEN 80® (SigmaAldrich, St. Louis, Mo.). Polysorbate 80 comprises approximately 60%-70%oleic acid with the remainder comprising primarily linoleic acids,palmitic acids, and stearic acids. Polysorbate 80 may be used in amountsranging from about 5 to 50%, and in certain embodiments, about 30% ofthe formulation total mass.

In various other embodiments, the non-ionic surfactant is selected fromone or more of glycerol and polyethylene glycol esters of long chainfatty acids, for example, lauroyl macrogol-32 glycerides and/or lauroylpolyoxyl-32 glycerides, commercially available as Gelucire, including,for example, Gelucire 44/11 and Gelucire 44/14. These surfactants may beused at concentrations greater than about 0.01%, and typically invarious amounts of about 0.01%-10.0%, 10.1%-20%, and 20.1%-30%.

In other embodiments, a lubricant is used. Any suitable lubricant may beused, such as for example lecithin. Lecithin may comprise a mixture ofphospholipids.

In additional embodiments, an antioxidant is used. Any suitableanti-oxidant may be used such as, for example and without limitationbutylated hydroxytoluene.

For example, in various embodiments, a pharmaceutical formulationcomprises about 20% to about 80% carrier by weight, about 0.1% to about5% lubricant by weight, and about 0.01% to about 0.1% antioxidant byweight.

The choice of excipient will, to a large extent, depend on factors suchas the particular mode of administration, the effect of the excipient onsolubility and stability, and the nature of the dosage form. Excipientsused in various embodiments may include colorants, flavoring agents,preservatives and taste-masking agents. Colorants, for example, maycomprise about 0.1% to about 2% by weight. Preservatives may comprisemethyl and propyl paraben, for example, in a ratio of about 10:1, and ata proportion of about 0.005% and 0.05% by weight.

As is with all oils, solubilizers, excipients and any other additivesused in the formulations described herein, each is to be non-toxic andpharmaceutically acceptable.

As referenced above, the formulations of the present disclosure aregenerally orally administered, typically via, for example, capsules suchas soft capsules. The present formulations can also be used to formtransdermal patches using standard technology known in the art.Solubilized formulations of the present invention can also be formulatedfor intraperitoneal administration using techniques well known in theart.

In accordance with various embodiments, formulations do not includepeanut oil. The lack of peanut oil obviates the risk posed to thosehaving peanut-based allergies.

According to various embodiments described herein, a 28-day or monthlyregimen of capsules can be packaged in a single kit (e.g., a blisterpack) having administration days identified to improve compliance andreduce associated symptoms, among others. One or more of the capsulesmay contain no estradiol, for example, and/or no progesterone. Capsulesthat comprise no estrogen or progesterone API may be referred to asplacebos. A blister pack can have a plurality of scores or perforationsseparating blister pack into 28 days. Each day may further comprise asingle blister or a plurality of blisters. In various embodiments, eachunit dose may contain micronized and/or partially solubilized, or fullysolubilized progesterone and/or solubilized estradiol in amounts as setforth herein above, although other dose ranges may be contemplated. Inaddition, kits having other configurations are also contemplated herein.For example, without limitation, kits having such blister packs maycontain any number of capsules.

Orally administered formulations of the present disclosure containingmicronized and/or partially solubilized, or fully solubilized,progesterone are also used for the treatment of endometrial hyperplasia,secondary amenorrhea and other disease states treated with supplementalprogesterone. Generally, progesterone-containing formulations describedherein are used to treat the effects of the administration ofsupplemental estrogen whether administered alone or in combination withsolubilized estradiol of the present disclosure or otherestrogen-containing formulations. In various other embodiments, acapsule containing formulations of the present disclosure, for example asoftgel capsule, may be applied in or around the vagina.

Formulations of the present disclosure containing solubilized estradiolare used to treat Estrogen-deficient States, including vasomotorsymptoms, for example, in relation to treatment of hypoestrogenismrelated symptoms including, for example and without limitation, hotflashes and night sweats (vasomotor symptoms), sleep disturbances, moodchanges, vulvo-vaginal atrophy, and osteoporosis and othernon-menopausal disease states treated with supplemental estrogen.

Formulations of the present disclosure containing solubilized estradiolmay be used to treat or prevent atrophic vaginitis or vulvo-vaginalatrophy. In various embodiments, a capsule, for example a softgelcapsule, may be applied in or around the vagina.

Additional objects of the present disclosure includes: providingincreased patient compliance secondary to ease of use; providingincreased physician adoption secondary to ease of use/instruction withless worry of side effects from inappropriate usage; providing decreasedside-effects from erroneous use (decreased irregular bleeding);providing better efficacy/control of symptoms secondary to appropriateuse; reducing the metabolic and vascular side effects of the commonlyused synthetic progestins when administered alone or in combination withan estrogen (norethindrone acetate, medroxyprogesterone acetate, etc.)including, for example, stroke, heart attacks, blood clots and breastcancer.

EXAMPLES Example 1 Estradiol Solubility

In various experiments, suitable solvents were determined for providingsufficient solubility to make 2 mg of estradiol in a 100 mg fill mass,with a desired goal of achieving ˜20 mg/g solubility for estradiol.Initial solubility experiments were done by mixing estradiol withvarious solvents, saturate the solution with the estradiol, equilibratefor at least 3 days and filter the un-dissolved particles and analyzingthe clear supernatant for the amount of estradiol dissolved by HPLC.

Estradiol solubility experiments were performed. From this list at leastone item (e.g. propylene glycol) is known to be unsuitable forencapsulation.

TABLE 1 Ingredient Solubility (mg/g) PEG 400 105*  Propylene Glycol 75*Polysorbate 80 36* TRANSCUTOL HP (Highly purified diethylene 141  glycolmonoethyl ether EP/NF) CAPMUL PG-8 (Propylene Glycol Monocaprylate) 31.2 *Literature reference -Salole, E. G. (1987) The PhysicochemicalProperties of Oestradiol, J Pharm and Biomed Analysis, 5, 635-640.

Example 2

It was desired to achieve 50 mg of progesterone suspended in a mediumthat can also solubilize 2 mg estradiol in a total capsule fill mass of200 mg. In order to achieve this formulation, the required solubility ofestradiol needs to be ˜10 mg/g. A total fill weight of 200 mg wasconsidered suitable for a size 5 oval soft gelatin capsule.

Additional solubility studies were performed to find solvent mixturesthat might possibly be more suitable for soft gelatin encapsulation.Solubility studies were conducted with CAPMUL PG-8 (Propylene GlycolMonocaprylate) and CAPMUL MCM (Medium Chain Mono- and Diglycerides) bymixing estradiol with various the solvent systems and as before byanalyzing for the amount of estradiol dissolved by HPLC afterfiltration. Results of these experiments are presented in Table 2. Itcan be seen from these results that mixtures containing MIGLYOL(caprylic/capric triglyceride):CAPMUL PG-8 (Propylene GlycolMonocaprylate) at 50%; and also CAPMUL MCM (Medium Chain Mono- andDiglycerides) alone or in combination with 20% Polysorbate 80 canachieve sufficient solubility to meet the target of 10 mg/g. CAPMUL PG-8(Propylene Glycol Monocaprylate) mixed with MIGLYOL (caprylic/caprictriglyceride) at the 15 and 30% level did not provide sufficientsolubility.

TABLE 2 Ingredient Solubility (mg/g) MIGLYOL (caprylic/caprictriglyceride):CAPMUL 4.40 PG-8 (Propylene Glycol Monocaprylate) (85:15)MIGLYOL (caprylic/capric triglyceride):CAPMUL 8.60 PG-8 (PropyleneGlycol Monocaprylate) (70:30) TRANSCUTOL (Diethylene glycolmonoethyl >12 ether):MIGLYOL 812 (Caprylic/Capric Triglyceride):CAPMULPG-8 (Propylene Glycol Monocaprylate) (5:65:28) TRANSCUTOL (Diethyleneglycol monoethyl >12 ether):MIGLYOL 812 (Caprylic/CapricTriglyceride):CAPMUL PG-8 (Propylene Glycol Monocaprylate) (5:47:47)MIGLYOL (caprylic/capric triglyceride):CAPMUL 14.0 PG-8 (PropyleneGlycol Monocaprylate) (50:50) CAPMUL MCM (Medium Chain Mono- and 19.8Diglycerides) Polysorbate 80:CAPMUL MCM (Medium Chain 15.0 Mono- andDiglycerides) (20:80)

Example 3

Additional studies were performed to assess the stability of estradiol(4-6 mg) in solvent mixtures, as reported in Table 3. MIGLYOL 812(Caprylic/Capric Triglyceride) with 4% TRANSCUTOL (Diethylene glycolmonoethyl ether) precipitated on Hot/Cold cycling after 96 hours, whileestradiol solubilized in MIGLYOL (caprylic/capric triglyceride):CAPMUL(a propylene glycol monocaprylate; propylene glycol monocaprate) blendsat 30 and 50% or in CAPMUL MCM (Medium Chain Mono- and Diglycerides)alone, did not precipitate under the same conditions for a minimum of 14days.

TABLE 3 Results Hot/Cold Formulation Estradiol mg/g Cycling TRANSCUTOL(Diethylene glycol 4 Crystallizes monoethyl ether):MIGLYOL after 96hours 812 (Caprylic/Capric Triglyceride) (4:96) MIGLYOL 812(Caprylic/Capric 6 Clear, after 14 days Triglyceride):CAPMUL PG-8(Propylene Glycol Monocaprylate) (70:30) MIGLYOL 812 (Caprylic/Capric 6Clear, after 14 days Triglyceride):CAPMUL PG-8 (Propylene GlycolMonocaprylate) (50:50) TRANSCUTOL (Diethylene glycol 6 Clear, after 14days monoethyl ether):MIGLYOL 812 (Caprylic/Capric Triglyceride):CAPMULPG-8 (Propylene Glycol Monocaprylate) (5:80:15) CAPMUL MCM (Medium 6Clear after 14 days Chain Mono- and Diglycerides)

12 mg estradiol solubilized in MIGLYOL (caprylic/caprictriglyceride):CAPMUL PG-8 (Propylene Glycol Monocaprylate) 50:50, CAPMULMCM(Medium Chain Mono- and Diglycerides), and in mixtures of TRANSCUTOL(Diethylene glycol monoethyl ether): MIGLYOL (caprylic/caprictriglyceride):CAPMUL PG-8 (Propylene Glycol Monocaprylate) are stableand do not precipitate for at least 12 days.

TABLE 4 Results Hot/Cold Formulation Estradiol mg/g Cycling MIGLYOL 812(Caprylic/Capric 12 Clear, after 12 days Triglyceride):CAPMUL PG-8(Propylene Glycol Monocaprylate) (50:50) TRANSCUTOL (Diethylene glycol12 Clear, after 12 days monoethyl ether):MIGLYOL 812 (Caprylic/CapricTriglyceride):CAPMUL PG-8 (Propylene Glycol Monocaprylate) (5:65:28)TRANSCUTOL (Diethylene glycol 12 Clear, after 12 days monoethylether):MIGLYOL 812 (Caprylic/Capric Triglyceride):CAPMUL PG-8 (PropyleneGlycol Monocaprylate) (5:47:47) CAPMUL MCM(Medium 12 Clear after 12 daysChain Mono- and Diglycerides)

Example 4

In addition to determining physical stability of the estradiol solutionsover time, it is necessary to determine if the fill material will bestable during the encapsulation process. One way to test thesepreparations is with the addition of water to the fill mass. As can beseen in Table 5, estradiol solutions at a concentration of 6 mg/g inPolyethylene Glycol 400 and CAPMUL MCM (Medium Chain Mono- andDiglycerides) are able to absorb a minimum of 7% water withoutrecrystallization, whereas the same concentration in MIGLYOL 812(Caprylic/Capric Triglyceride):CAPMUL PG-8 (Propylene GlycolMonocaprylate) (75:25) precipitates.

Estradiol solutions at a concentration of 12 mg/g in Polyethylene Glycol400 and CAPMUL MCM (Medium Chain Mono- and Diglycerides) are able toabsorb a minimum of 7% water without recrystallization. All CAPMUL PG-8(Propylene Glycol Monocaprylate) containing formulations turned hazy onthe addition of water. However, it should be noted that estradiolrecrystallization was not observed, and the addition of water to CAPMULPG-8 (Propylene Glycol Monocaprylate) alone (without any estradiol) alsoturns hazy on the addition of water.

TABLE 5 Results after Estradiol addition of 7% Formulation mg/g waterMIGLYOL 812 (Caprylic/Capric 6 Precipitated Triglyceride):CAPMUL PG-8(Propylene Glycol Monocaprylate) (75:25) MIGLYOL 812 (Caprylic/Capric 12Hazy Triglyceride):CAPMUL PG-8 (Propylene Glycol Monocaprylate) (50:50)TRANSCUTOL (Diethylene glycol 12 Hazy monoethyl ether):MIGLYOL 812(Caprylic/Capric Triglyceride):CAPMUL PG-8 (Propylene GlycolMonocaprylate) (5:65:28) CAPMUL MCM (Medium Chain Mono- 12 Clear andDiglycerides) TRANSCUTOL (Diethylene glycol 12 Hazy monoethylether):MIGLYOL 812 (Caprylic/Capric Triglyceride):CAPMUL PG-8 (PropyleneGlycol Monocaprylate) (5:47:47) Polyethylene Glycol 400 12 clear

Example 5

In an exemplary embodiment, a capsule is provided containing a fillmaterial comprising:

TABLE 6 Ingredient Mg/Capsule Estradiol Hemihydrate 2.00 Mono-, di- ortriglyceride (Miglyol qs 812) Diethylene Glycol Monoethylether 65.00(TRANSCUTOL HP (Highly purified diethylene glycol monoethyl etherEP/NF)) Liquid lecithin 1.63 Butylated Hydroxytoluene 0.13 Total FillWeight 325

Example 6

In an exemplary embodiment, a capsule is provided containing a fillmaterial comprising:

TABLE 7 Ingredient Mg/Capsule Estradiol Hemihydrate 2.00Monoglycerides/diglycerides/triglycerides of qs caprylic/capric acid(CAPMUL MCM (Medium Chain Mono- and Diglycerides)) Liquid lecithin 1.63Polysorbate 80 97.5 Total Fill Weight 325

In an exemplary embodiment, a capsule is provided containing a fillmaterial comprising:

TABLE 8 Mg/ Amount/ Ingredient Capsule % w/w Batch Estradiol Hemihydrate2.03 0.62 20.2 g Monoglycerides/diglycerides/triglycerides 322.97 99.383.23 kg of caprylic/capric acid (CAPMUL MCM (Medium Chain Mono- andDiglycerides)) Total 100 3.25 kg

The above formulation is prepared as follows: estradiol is added toCAPMUL MCM (Medium Chain Mono- and Diglycerides) and mixed untildissolved.

Example 7 Progesterone Solubility

In various embodiments, both estradiol and progesterone may be dissolvedin a solvent. In various embodiments, the solubility of both estradioland progesterone will be such that a therapeutically effective dose maybe obtained in a reasonably sized mass, generally considered to bebetween 1 mg and 1200 mg, preferably suitable for encapsulation in asize 3 to 22 oval or oblong capsule. For example, in variousembodiments, 50 mg to 100 mg of progesterone may be dissolved in avolume of solvent; i.e., the solubility would be 50 mg to 100 mg percapsule. MIGLYOL (caprylic/capric triglyceride) was attempted, and whileit can be considered a good carrier for progesterone, it alone did notprovide a desirable level of solubilization of estradiol (e.g.,solubility of 12 mg/g may be desirable in various embodiments). Thus,MIGLYOL (caprylic/capric triglyceride) may be used in embodimentscomprising a suspension of progesterone, though MIGLYOL (caprylic/caprictriglyceride), standing alone, is not desirable for use in embodimentshaving fully solubilized progesterone and/or estradiol.

As can be seen in Table 9, the solubility of progesterone in CAPMUL MCM(Medium Chain Mono- and Diglycerides) is ˜73 mg/g. Therefore, bysuspending 200 mg progesterone in 400 mg of solvent, part of the dose(˜14%) is already dissolved and the remaining is still a suspension. Insome aspects and embodiments, it is desired to minimize the partialsolubility of progesterone in the formulation in order to minimize thepossibility of recrystalization.

Based on 73 mg/g solubility, the capsule size required to make a capsuleof 50 mg solubilized progesterone would be 685 mg. Therefore, it wasshown that it would be feasible to make a 50 mg progesterone and 2 mgestradiol solubilized formulation. MIGLYOL (caprylic/caprictriglyceride) had the lowest solubility, but that solvent is unable todissolve the estradiol, therefore under further experiments, it wasdecided to proceed with the second lowest or CAPMUL MCM (Medium ChainMono- and Diglycerides). It has also been found that 2 mg of estradiolmay also be dissolved in 685 mg of CAPMUL MCM (Medium Chain Mono- andDiglycerides).

TABLE 9 Progesterone Solubility Ingredient (mg/g) CAPMUL MCM (Medium73.4 Chain Mono- and Diglycerides) CAPMUL PG-8 (Propylene 95 GlycolMonocaprylate) MIGLYOL 812 (Caprylic/ 27.8 Capric Triglyceride)

In addition, it has been found that the solubility of progesterone in asolvent of CAPMUL MCM (Medium Chain Mono- and Diglycerides) incombination with Gelucire 44/14 in a 9:1 ratio increases the solubilityto approximately 86 mg/g. Therefore, in various embodiments,progesterone and/or estradiol may be dissolved in a CAPMUL MCM (MediumChain Mono- and Diglycerides) and Gelucire 44/14 system, wherein theratio of CAPMUL MCM (Medium Chain Mono- and Diglycerides) to GELUCIRE44/14 (Lauroyl macrogol-32 glycerides EP Lauroyl polyoxyl-32 glyceridesNF Lauroyl polyoxylglycerides (USA FDA IIG)) is 9:1.

TABLE 10 Progesterone Solubility Ingredient (mg/g) CAPMUL MCM (MediumChain Mono- 86.4 and Diglycerides):GELUCIRE 44/14 (Lauroyl macrogol-32glycerides EP Lauroyl polyoxyl-32 glycerides NF Lauroylpolyoxylglycerides (USA FDA IIG)) (9:1) CAPMUL MCM (Medium Chain Mono-70.5 and Diglycerides) GELUCIRE 44/14 (Lauroyl macrogol-32 glycerides EPLauroyl polyoxyl-32 glycerides NF Lauroyl polyoxylglycerides (USA FDAIIG)) (7:3) CAPMUL MCM (Medium Chain Mono- 57.4 andDiglycerides):GELUCIRE 44/14 (Lauroyl macrogol-32 glycerides EP Lauroylpolyoxyl-32 glycerides NF Lauroyl polyoxylglycerides (USA FDA IIG))(6:4)

Example 7

In an exemplary embodiment, a capsule is provided containing a fillmaterial having fully solubilized progesterone and estradiol comprising:

TABLE 11 Mass Qty/Capsule Ingredient (mg) % w/w (mg) Progesterone, USP,micronized 50.00 7.14 50.00 Estradiol Hemihydrate, USP 2.03 0.29 2.03CAPMUL MCM (Medium Chain 82.57 577.97 Mono- and Diglycerides), NFGELUCIRE 44/14 (Lauroyl macrogol- 10.0 70.00 32 glycerides EP Lauroylpolyoxyl-32 glycerides NF Lauroyl polyoxylglycerides (USA FDA IIG)), NFTOTAL 100.00 700.00

A capsule such as that shown in TABLE 11 may be manufactured in anysuitable manner. For the purposes of this Example, mixing may befacilitated by an impellor, agitator, or other suitable means. Also forthe purposes of this Example, heating and/or mixing may be performedunder an inert or relatively inert gas atmosphere, such as nitrogen gasN2. Mixing and/or heating for the purposes of this Example may beperformed in any suitable vessel, such as a stainless steel vessel.

For example, CAPMUL MCM (Medium Chain Mono- and Diglycerides) may beheated to between 30° C. to 50° C., more preferably from 35° C. to 45°C., and more preferably to 40° C.+/−2° C. GELUCIRE 44/14 (Lauroylmacrogol-32 glycerides EP Lauroyl polyoxyl-32 glycerides NF Lauroylpolyoxylglycerides (USA FDA IIG)) may be added to the CAPMUL MCM (MediumChain Mono- and Diglycerides) and mixed until dissolved. The additionmay occur all at once or may occur gradually over a period of time. Heatmay continue to be applied during the mixing of the Gelucire 44/14 andthe CAPMUL MCM (Medium Chain Mono- and Diglycerides).

Heat may be removed from the GELUCIRE 44/14 (Lauroyl macrogol-32glycerides EP Lauroyl polyoxyl-32 glycerides NF Lauroylpolyoxylglycerides (USA FDA IIG)) and CAPMUL MCM (Medium Chain Mono- andDiglycerides) mixture. Estradiol Hemihydrate may be added to themixture. The addition may occur all at once or may occur gradually overa period of time. Micronized progesterone may then be added to theGelucire 44/14, CAPMUL MCM (Medium Chain Mono- and Diglycerides) andEstradiol Hemihydrate mixture until dissolved. The addition may occurall at once or may occur gradually over a period of time.

Example 8

In an exemplary embodiment, a capsule is provided containing a fillmaterial having suspended progesterone comprising:

TABLE 12 Ingredient mg/Capsule % Function Micronized 200.00 30.77 ActiveProgesterone Medium Chain qs qs Carrier Triglyceride (MIGLYOL 812(Caprylic/ Capric Triglyceride) or equivalent) Lecithin Liquid 1.63 0.25Lubricant/ Emulsifier Butylated 0.13 0.02 Antioxidant Hydroxytoluene(also referred to as “BHT”)

The above formulation is prepared as follows: MIGLYOL (caprylic/caprictriglyceride) is heated to about 45° C. GELUCIRE 44/14 (Lauroylmacrogol-32 glycerides EP Lauroyl polyoxyl-32 glycerides NF Lauroylpolyoxylglycerides (USA FDA IIG)) is added and mixed until dissolved.BHT is added and mixed until dissolved. Progesterone is suspended andpassed through a colloid mill. The resultant fill mass can be used forencapsulation.

In an exemplary embodiment, a capsule is provided containing a fillmaterial having partially solubilized progesterone comprising:

TABLE 13 Qty/ Qty/ Amount/ Capsule Capsule Batch Ingredient (mg) % w/w(mg) (kg) Micronized 200.00 33.33 Active 2.0 Progesterone, USPMonoglycerides/ 394.0 65.67 Carrier 3.94 diglycerides/triglycerides ofcaprylic/capric acid (CAPMUL MCM (Medium Chain Mono- and Diglycerides))Lauroyl polyoxyl-32-  6.0 1 Lubricant/ 0.06 glycerides Emulsifier(GELUCIRE 44/14 (Lauroyl macrogol-32 glycerides EP Lauroyl polyoxyl-32glycerides NF Lauroyl polyoxylglycerides (USA FDA IIG)) or equivalent)Total 600.00 mg 100 6.0 kg

For suspensions of progesterone and partially solubilized progesterone,GELUCIRE 44/14 may be added at 1% to 2% w/w to increase viscosity. Theabove formulation is prepared as follows: CAPMUL MCM (Medium Chain Mono-and Diglycerides) is heated to about 65° C. GELUCIRE 44/14 (Lauroylmacrogol-32 glycerides EP Lauroyl polyoxyl-32 glycerides NF Lauroylpolyoxylglycerides (USA FDA IIG)) is added and mixed until dissolved.Heat is removed. Progesterone is added and the mixture is passed througha colloid mill. The resultant fill mass can be used for encapsulation.

Example 9

In an exemplary embodiment, a capsule is provided containing a fillmaterial having suspended progesterone comprising:

TABLE 14 Ingredient % mg/Capsule Function Micronized Progesterone 30.77200.00 Active Medium Chain Triglyceride 65.93 428.55 Carrier (MIGLYOL812 (Caprylic/ Capric Triglyceride) or equivalent) Lauroylpolyoxyl-32-glycerides 3.00 19.50 Suspending (GELUCIRE 44/14 (LauroylAgent macrogol-32 glycerides EP Lauroyl polyoxyl-32 glycerides NFLauroyl polyoxylglycerides (USA FDA IIG)) or equivalent) ButylatedHydroxytoluene 0.03 1.95 Antioxidant Total 100 650

In various embodiments, amounts of MIGLYOL (caprylic/caprictriglyceride) may be present in a range from about 35-95% by weight;GELUCIRE 44/14 (Lauroyl macrogol-32 glycerides EP Lauroyl polyoxyl-32glycerides NF Lauroyl polyoxylglycerides (USA FDA IIG)) from about0.5-30% by weight; and BHT from about 0.01-0.1% by weight.

Example 10

For the purposes of this Example, a particle size analysis is conductedby using the Beckman Device. A sample API comprising micronizedprogesterone in accordance with various embodiments is provided foranalysis.

Approximately 0.01 g of a sample API in accordance with variousembodiments was combined with Coulter 1B and 10 mL of deionized water.Sonication was performed for 15 seconds. The Beckman Device, equippedwith a ULM, performed analysis for 90 seconds. The Beckman Device wasconfigured to use the Fraunhofer optical model. The Beckman Deviceyielded that the sample has an X50 of 4.279 μm, an X75 of 7.442 μm, andan X25 of 1.590 μm. The Beckman Device also yielded that the meanparticle size is 4.975 μm, the median particle size is 4.279 μm, themode particle size is 6.453 μm, and the standard deviation is 3.956 μm.A graph of the particle distribution obtained is shown in FIG. 4.

Example 11

A formulation sample having approximately 200 mg of micronizedprogesterone and 2 mg of estradiol was dispersed with oil. The BeckmanDevice, equipped with a MLM, performed analysis for 60 seconds. TheBeckman Device was configured to use the Fraunhofer optical model. TheBeckman Device yielded that the sample has an X50 of 11.0 μm, an X75 of17.3 μm, and an X25 of 5.3 μm. The Beckman Device also yielded that themean particle size is 11.8 μm, the median particle size is 11.04 μm, themode particle size is 13.6 μm, and the standard deviation is 7.8 μm.

Example 12

In order to increase the solubility of progesterone in the finalsolution, GELUCIRE 44/14 (Lauroyl macrogol-32 glycerides EP Lauroylpolyoxyl-32 glycerides NF Lauroyl polyoxylglycerides (USA FDA IIG)) wasadded at about 10% w/w.

TABLE 15 Quantitative Formula: Batch Size 10,000 capsules Label Qty/Amount/ Item Claim Capsule Batch No. Ingredient(s) (mg) % w/w (mg)(kg) 1. Progesterone, USP, 50.00 7.14 50.00 0.50 micronized 2. EstradiolHemihydrate, 2.03 0.29 2.03 0.02 USP 3. CAPMUL MCM 82.57 577.97 5.78(Medium Chain Mono- and Diglycerides), NF 4. GELUCIRE 44/14 10.0 70.000.70 (Lauroyl macrogol-32 glycerides EP Lauroyl polyoxyl-32 glyceridesNF Lauroyl polyoxylglycerides (USA FDA IIG)) Gelucire 44/14, NF Total:100.00 700.00 7.00

An example of the final formulation is provided in Table 15. Themanufacturing process is as follows. CAPMUL MCM (Medium Chain Mono- andDiglycerides) is heated to 40° C. GELUCIRE 44/14 (Lauroyl macrogol-32glycerides EP Lauroyl polyoxyl-32 glycerides NF Lauroylpolyoxylglycerides (USA FDA IIG)) is added and mixed until dissolved.Heat is removed. Estradiol is added and mixed until dissolved.Micronized progesterone is then added and mixed until dissolved.

Example 13

In an exemplary embodiment, a capsule is provided containing a fillmaterial having fully solubilized estradiol and partially solubilizedprogesterone comprising:

TABLE 16 Label Qty/ Amount/ Item Claim Capsule Batch No. Ingredient(s)(mg) % w/w (mg) (g) 1. Progesterone, USP, 50.00 25.000  50.00 500.00micronized 2. Estradiol Hemihydrate 0.25 0.129  0.26 2.58 3. CAPMUL MCM73.371 146.74 1467.42 (Medium Chain Mono- and Diglycerides), NF 4.GELUCIRE 44/14 1.500  3.00 30.00 (Lauroyl macrogol-32 glycerides EPLauroyl polyoxyl-32 glycerides NF Lauroyl polyoxylglycerides (USA FDAIIG)), NF Total: 100.000 200.00 mg 2000.00

The manufacturing process is as follows. CAPMUL MCM (Medium Chain Mono-and Diglycerides) is heated to 65° C. GELUCIRE 44/14 (Lauroylmacrogol-32 glycerides EP Lauroyl polyoxyl-32 glycerides NF Lauroylpolyoxylglycerides (USA FDA IIG)) is added and mixed until dissolved.Heat is removed. Estradiol is added and mixed until dissolved.Micronized progesterone is then added and dispersed. The mixture is thenpassed through a colloid mill. The resultant fill mass can be used forencapsulation.

Example 14

In an exemplary embodiment, a capsule is provided containing a fillmaterial having fully solubilized estradiol and partially solubilizedprogesterone comprising:

TABLE 17 Label Qty/ Amount/ Item Claim Capsule Batch No. Ingredient(s)(mg) % w/w (mg) (g) 1. Progesterone, USP, 200.00 33.33 200.0 2000.0micronized 2. Estradiol Hemihydrate 2.00 0.35  2.07 20.7 3. CAPMUL MCM65.32 391.93 3919.3 (Medium Chain Mono- and Diglycerides), NF 4.GELUCIRE 44/14 1.00  6.0 60.0 (Lauroyl macrogol-32 glycerides EP Lauroylpolyoxyl-32 glycerides NF Lauroyl polyoxylglycerides (USA FDA IIG)), NFTotal: 100.00 600.0 mg 6000.0

The manufacturing process is as follows. CAPMUL MCM (Medium Chain Mono-and Diglycerides) is heated to 65° C. Gelucire 44/14 is added and mixeduntil dissolved. Heat is removed. Estradiol is added and mixed untildissolved. Micronized progesterone is then added and dispersed. Themixture is then passed through a colloid mill. The resultant fill masscan be used for encapsulation.

Example 15

Progesterone and Estradiol Combination Study Under Fed Conditions.

This following study protocol was used to establish bio-availability andbio-equivalence parameters for a combination product of the presentdisclosure comprising progesterone (200 mg) and estradiol (2.0 mg) asprepared via the process described in Example 14 and compared to 200 mgof PROMETRIUM (progesterone, USP) (Catalent Pharmaceuticals, St.Petersburg, Fla. (and 2.0 mg of ESTRACE (estradiol vaginal cream, USP,0.01%) (Bristol-Myers Squibb Co. Princeton, N.J.), administered totwenty-four (24) normal healthy, adult human post-menopausal femalesubjects under fed conditions.

The Study Design: An open-label, balanced, randomized, two-treatment,two-period, two-sequence, single-dose, two-way crossover.

The subjects were housed in the clinical facility from at least 11.00hours pre-dose to at least 48.00 hours post-dose in each period, with awashout period of at least 14 days between the successive dosing days.

Subjects were fasted for at least about 10.00 hours before being serveda high-fat, high-calorie breakfast, followed by dosing, then followed bya 04.00 hour, post-dose additional period of fasting.

Standard meals were provided at about 04.00, 09.00, 13.00, 25.00, 29.00,34.00 and 38.00 hours post-dose, respectively.

Water was restricted at least about 01 hour prior to dosing until about01 hour post-dose (except for water given during dosing). At othertimes, drinking water was provided ad libitum.

Subjects were instructed to abstain from consuming caffeine and/orxanthine containing products (i.e. coffee, tea, chocolate, andcaffeine-containing sodas, colas, etc.) for at least about 24.00 hoursprior to dosing and throughout the study, grapefruit and\or its juiceand poppy containing foods for at least about 48.00 hours prior todosing and throughout the study.

Subjects remained seated upright for about the first 04.00 hourspost-dose and only necessary movements were allowed during this period.Thereafter subjects were allowed to ambulate freely during the remainingpart of the study. Subjects were not allowed to lie down (except asdirected by the physician secondary to adverse events) duringrestriction period.

Subjects were instructed not to take any prescription medications within14 days prior to study check in and throughout the study. Subjects wereinstructed not to take any over the counter medicinal products, herbalmedications, etc. within 7 days prior to study check-in and throughoutthe study.

After overnight fasting of at least about 10.00 hours, a high-fathigh-calorie breakfast was served about 30 minutes prior toadministration of investigational product(s). All subjects were requiredto consume their entire breakfast within about 30 minutes of it beingserved, a single dose of either test product (T) of Progesterone 200 mg& Estradiol 2 mg tablets or the reference product (R) PROMETRIUM(progesterone, USP) soft gel Capsule 200 mg and ESTRACE (estradiolvaginal cream, USP, 0.01%) (Estradiol) Tablets 2 mg (according to therandomization schedule) were administered with about 240 mL of waterunder fed condition, at ambient temperature in each period in sittingposture. A thorough mouth check was done to assess the compliance todosing.

All dosed study subjects were assessed for laboratory tests at the endof the study or as applicable.

In each period, twenty-three (23) blood samples were collected. Thepre-dose (10 mL) blood samples at −01.00, −00.50, 00.00 hours and thepost-dose blood samples (08 mL each) were collected at 00.25, 00.50,00.67, 00.83, 01.00, 01.33, 01.67, 02.00, 02.50, 03.00, 04.00, 05.00,06.00, 07.00, 08.00, 10.00, 12.00, 18.00, 24.00 and 48.00 hours inlabeled K2EDTA—vacutainers via an indwelling cannula placed in one ofthe forearm veins of the subjects. Each intravenous indwelling cannulawas kept in situ as long as possible by injecting about 0.5 mL of 10IU/mL of heparin in normal saline solution to maintain the cannula forcollection of the post-dose samples. In such cases blood samples werecollected after discarding the first 0.5 mL of heparin containing blood.Each cannula was removed after the 24.00 hour sample was drawn orearlier or if blocked.

At the end of the study, the samples were transferred to thebio-analytical facility in a box containing sufficient dry ice tomaintain the integrity of the samples. These samples were stored at atemperature of −70° C.±20° C. in the bio-analytical facility untilanalysis.

Progesterone (Corrected and Uncorrected) and Estradiol (unconjugated)and estrone (total) in plasma samples is assayed using a validatedLC-MS/MS method.

Fasted studies using this protocol were also conducted. However, ratherthan the high-fat meal prior to administration of the test and referencedrug, each subject fasted for a period of at least twelve (12) hoursprior to dose administration.

Example 16

Method of manufacture in accordance with various embodiments are shownin FIGS. 1-3. With reference to FIG. 1, method of fill material 100 isshown. Step 102 comprises heating an oily vehicle carrier to 40° C.±5°C. Heating may be accomplished through any suitable means. The heatingmay be performed in any suitable vessel, such as a stainless steelvessel. The oily vehicle may be any oily vehicle described herein, forexample, CAPMUL MCM (Medium Chain Mono- and Diglycerides).

Step 104 comprises mixing Gelucire 44/14 with the oily vehicle. Mixingmay be facilitated by an impellor, agitator, or other suitable means.Step 102 may be performed under an inert or relatively inert gasatmosphere, such as nitrogen gas N₂. Mixing may be performed in anysuitable vessel, such as a stainless steel vessel.

Step 106 comprises mixing estradiol into the mixture of the oily vehicleand Gelucire 44/14. Mixing may occur in a steel tank or vat. Mixing maybe facilitated by an impellor, agitator, or other suitable means. Step106 may be performed under an inert or relatively inert gas atmosphere,such as nitrogen gas N₂.

Step 108 comprises cooling to room temperature. Cooling may be allowedto occur without intervention or cooling may be aided by application ofa cooling system.

Step 110 comprises mixing micronized progesterone into the mixture ofoily vehicle, estradiol and Gelucire 44/14. Mixing may occur in a steeltank or vat. Mixing may be facilitated by an impellor, agitator, orother suitable means. Step 110 may be performed under an inert orrelatively inert gas atmosphere, such as nitrogen gas N₂. Step 112comprises degasing. The resulting mixture from step 112 may comprise afill material suitable for production into a softgel capsule.

With reference to FIG. 2, softgel capsule, i.e. gel mass, production 200is shown. Step 202 comprises mixing glyercin with water. The water usedin step 202 may be purified by any suitable means, such as reverseosmosis, ozonation, filtration (e.g., through a carbon column) or thelike. Mixing may be facilitated by an impellor, agitator, or othersuitable means. Step 202 may be performed under an inert or relativelyinert gas atmosphere, such as nitrogen gas N₂. Heating may be performeduntil the temperature reaches 80° C.±5° C.

Step 204 comprises the addition of gelatin to the glycerin watermixture. Mixing may be facilitated by an impellor, agitator, or othersuitable means. Step 204 may be performed under an inert or relativelyinert gas atmosphere, such as nitrogen gas N₂. A vacuum may be drawn instep 204 to de-aerate.

Step 206 comprises addition of a coloring agent such as a dye. Acoloring agent may comprise products sold under the trademark OPATINT orother suitable agent. Step 206 may be performed under an inert orrelatively inert gas atmosphere, such as nitrogen gas N₂. Step 208comprises degasing. The resulting mixture from step 208 may comprise agel capsule material suitable for use as a gel capsule in production ofa softgel capsule.

With reference to FIG. 3, softgel capsule assembly process 300 is shown.Step 302 comprises heating the fill material. The fill material may beheated to any suitable temperature. In various embodiments, the fillmaterial is heated to 30° C.+/−3° C. Fill material maybe heated in afill hopper. A fill hopper may comprise a device configured to hold avolume of the fill material and/or to dispense the fill material incontrolled volumes. Step 304 comprises filling a gel mass. A gel massmay be taken from the gel capsule material produced in step 208 of FIG.2. Filling may be performed by injecting, placing, or otherwisedisposing the fill material within a volume defined by the gel capsulematerial. The filling may occur in an encapsulator. The spreader boxesmay be a temperature of 55° C.+/−10° C. The wedge temperature may be 38°C.+/−3° C. The drum cooling temperature may be 4° C.+/−2° C. Theencapsulator may be lubricated using MIGLYOL 812 (Caprylic/CapricTriglyceride) or other suitable lubricant. Step 304 thus produces one ormore softgel capsules. Filling may comprise producing a ribbon ofthickness 0.85 mm±0.05 mm using spreader box knobs. The fill materialmay be injected into the gel to produce a fill weight having targetweight±5% (i.e., 650±33 mg and 325±16.3 mg).

Step 304 comprises filling a gel mass. A gel mass may be taken from thegel capsule material produced in step 208 of FIG. 2. Filling may beperformed by injecting, placing, or otherwise disposing the fillmaterial within a volume defined by the gel capsule material. Thefilling may occur in an encapsulator. The spreader boxes may be atemperature of 55° C.+/−10° C. The wedge temperature may be 38° C.+/−3°C. The drum cooling temperature may be 4° C.+/−2° C. The encapsulatormay be lubricated using MIGLYOL 812 or other suitable lubricant. Step304 thus produces one or more softgel capsules. Filling may compriseproducing a ribbon of thickness 0.85 mm±0.05 mm using spreader boxknobs. The fill material may be injected into the gel to produce a fillweight having target weight±5% (i.e., 650±33 mg and 325±16.3 mg).

Step 306 comprises drying the softgel capsules. Drying may be performedin a tumble dryer, tray dryer, or combinations thereof. For example,drying may be performed in a tumble drying basket for between about 10minutes and about 120 minutes. Drying may continue in a drying room forabout 24 hours to about 72 hours. Step 308 may comprise inspectionand/or polishing. Polishing may be performed with isopropyl alcohol.Step 310 may comprise packaging. Packaging may be accomplished throughany suitable means. Packaging may comprise packing softgel capsules intoa blister pack, bottle, box, pouch, or other acceptable packaging.

We claim:
 1. A pharmaceutical composition comprising: a solubilizingagent comprising: mono- and diglycerides of capric and caprylic acid;and at least one of lauroyl macrogol-32 glycerides EP, lauroylpolyoxyl-32 glycerides NF, or lauroyl polyoxylglycerides; progesterone;and estradiol, the estradiol being at least about 90% solubilized in thesolubilizing agent; wherein the estradiol and the progesterone arepresent in the solubilizing agent, and the estradiol and suspendedprogesterone are uniformly dispersed.
 2. The pharmaceutical compositionof claim 1, wherein the ratio of progesterone to estradiol is from about24:1 to about 200:1.
 3. The pharmaceutical composition of claim 2,wherein the ratio of progesterone to estradiol comprises one of: about24:1, about 25:1, about 96:1, about 100:1, about 192:1, and about 200:1.4. The pharmaceutical composition of claim 1, wherein the progesteroneis between about 7.14% w/w and about 33.33% w/w of the pharmaceuticalcomposition.
 5. The pharmaceutical composition of claim 1, wherein theestradiol is between about 0.12% w/w and about 0.35% w/w of thepharmaceutical composition.
 6. The pharmaceutical composition of claim1, wherein the composition is encapsulated in a gelatin capsule; andwherein each gelatin capsule comprises from about 25 mg to about 200 mgof progesterone and from about 0.125 mg to about 2.00 mg of estradiol.7. The pharmaceutical composition of claim 1, wherein the estradiol isat least 90% solubilized in the solubilizing agent.
 8. A pharmaceuticalcomposition comprising: a solubilizing agent comprising: monoglyceridesand diglycerides of caprylic and capric acid; and a polyethylene glycolglyceride; progesterone; and estradiol, the estradiol being at leastabout 90% solubilized in the solubilizing agent; wherein the estradioland the progesterone are present in the solubilizing agent, and theestradiol and progesterone are uniformly dispersed.
 9. Thepharmaceutical composition of claim 8, wherein the ratio of progesteroneto estradiol is from about 24:1 to about 200:1.
 10. The pharmaceuticalcomposition of claim 9, wherein the ratio of progesterone to estradiolcomprises one of: about 24:1, about 25:1, about 96:1, about 100:1, about192:1 and about 200:1.
 11. The pharmaceutical composition of claim 8,wherein the progesterone is between about 7.14% w/w and about 33.33% w/wof the pharmaceutical composition.
 12. The pharmaceutical composition ofclaim 8, wherein the estradiol is between about 0.12% w/w and about0.35% w/w of the pharmaceutical composition.
 13. The pharmaceuticalcomposition of claim 8, wherein the composition is encapsulated in agelatin capsule; and wherein each gelatin capsule comprises from about25 mg to about 200 mg of progesterone and from about 0.125 mg to about2.00 mg of estradiol.
 14. The pharmaceutical composition of claim 8,wherein the estradiol is at least 90% solubilized in the solubilizingagent.
 15. A pharmaceutical composition comprising: a solubilizing agentcomprising: mono- and diglycerides of capric and caprylic acid; and atleast one of lauroyl macrogol-32 glycerides, lauroyl polyoxyl-32glycerides, and lauroyl polyoxylglycerides; progesterone; and estradiol,the estradiol being at least about 90% solubilized in the solubilizingagent; wherein the estradiol and the suspended progesterone are presentin the solubilizing agent, and the estradiol and suspended progesteroneare uniformly.
 16. The pharmaceutical composition of claim 15, whereinthe ratio of progesterone to estradiol is from about 24:1 to about200:1.
 17. The pharmaceutical composition of claim 15, wherein the ratioof progesterone to estradiol comprises one of: about 24:1, about 25:1,about 96:1, about 100:1, about 192:1 and about 200:1.
 18. Thepharmaceutical composition of claim 15, wherein the progesterone isbetween about 7.14% w/w and about 33.33% w/w of the pharmaceuticalcomposition.
 19. The pharmaceutical composition of claim 15, wherein theestradiol is between about 0.12% w/w and about 0.35% w/w of thepharmaceutical composition.
 20. The pharmaceutical composition of claim15, wherein the composition is encapsulated in a gelatin capsule; andwherein each gelatin capsule comprises from about 25 mg to about 200 mgof progesterone and from about 0.125 mg to about 2.00 mg of estradiol.